IL-8 anticorps

N° du produit ABIN2689785

Détail du produit anti-IL-8 anticorps

Antigène: IL-8 (IL8) (Interleukin 8 (IL8))

Reactivité: Humain

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp IL-8 est non-conjugé

Application: Immunocytochemistry (ICC)

Marque: BD Pharmingen™

Attributs du produit: The G265-8 antibody reacts with both the 72 and 77 amino acid forms of human interleukin-8 (IL-8). The immunogen used to produce the G265-8 hybridoma was E. coli-expressed recombinant human IL-8. PBMC were isolated from human peripheral blood by density gradient centrifugation and were cultured overnight at 37 °C with LPS (Sigma No. L-8274, 1 μg / ml) in the presence of GolgiStop™ (Cat. No. 554724). The activated cells were harvested and the level of IL-8 producing cells was detected by immunocytochemistry using a three-step staining procedure that employs a Biotin Goat anti-mouse IgG secondary antibody (Cat. No. 550337) and Streptavdin-horseradish peroxidase (Cat. No. 550946) (Nomarski optics, original magnification 400X).

BD Pharmingen™ Purified Mouse Anti-Human IL-8 - Purified - Clone G265-8 - Isotype Mouse IgG2b - Reactivity Hu - 0.25 mg

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogène: Recombinant Human IL-8

Clone: G265

Isotype: IgG2b

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Procédure de l'essai:

FOR IMMUNOCYTOCHEMICAL STAINING OF SINGLE-CELL PREPARATIONS This procedure describes the immunoenzymatic technique of staining cytokine within individual cells that are immobilized on microscopic slides via adherence (adherent slides) or centrifugation (cytospins). ADHESION SLIDES 1. Harvest cells and wash them twice in PBS using centrifugation (400 x g for 5 min) to remove residual protein. 2. Adjust the cell concentration at 4-5 x 10e6 cells/mL in PBS. 3. Place 20 μL of the cell suspension in each well of the adhesion slides and let them adhere at room temperature (RT) for 20 min. Please note that the slides should be washed in PBS at RT for 5 min before transferring the cells. 4. Fix cells on slides using fixation buffer for 15 min at RT. 5. Wash slides 2X in PBS with 5 min incubations. 6. Block slides with PBS supplemented with 1 % (w/v) BSA (Sigma, Cat. No. A43-78) for 30 min at RT or 10 min at 37 °C. 7. Wash slides 2X in PBS and proceed with staining or air dry them and store them at -80 °C for future use. 8. Incubate slides with 20 il of 1 % goat serum and PBS with 0.1 % (w/v) saponin for 30 min at RT. 9. Wash slides 2X with PBS with 5 min incubations. 10. Block endogenous peroxidase activity with Endogenous Peroxidase Blocking Buffer (20 μL/well) for 10 min at RT. 11. Wash 2X in PBS with 5 min incubations. 12. Incubate each well with Avidin (20 μL/well) for 15 min. 13. Wash 2X in PBS with 5 min incubations. 14. Incubate each well with Biotin (20 μL/well) for 15 min. 15. Wash 2X in PBS with 5 min incubations. 16. Incubate each well for 1 hr at RT with 20 μL of purified cytokine-specific antibody or appropriate immunoglobulin isotype control diluted in Antibody Diluent for IHC supplemented with saponin. 17. Wash slides 2X in PBS with 5 min incubations. 18. Incubate each well with 20 μL of a biotinylated secondary antibody diluted in Antibody Diluent for IHC for 30 min at RT. 19. Wash 2X in PBS with 5 min incubations. 20. Apply 20 μL of Streptavidin-HRP (BD Cat. No. 550946) to each well on slides and incubate for 30 min at RT. 21. Wash slides 2X with PBS with 5 minutes incubations. 22. Incubate with DAB Substrate as directed, (BD Cat. No. 550880) for less than 5 min at RT. 23. Stop the development of the color reaction by washing with PBS. 24. The slides are subsequently mounted in short-term storage mounting medium. CYTOSPINS 1. Assemble the Cytospin's sample chamber (e.g. Cytospin 3, Shandon, UK or comparable centrifuge), filter card, slide and cytospin racks according to manufacturer's specifications. 2. Load 40 μL of approximately 1 x 10e6 cells to each sample chamber. 3. Spin slides at 600 rpm for 2 min. 4. Take slides out of the cytospin rack and place them on a staining rack. 5. For fixation and staining please follow the steps 4 through 24 specified above for staining cells on adhesion slides.

Restrictions: For Research Use only

Stockage

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Agent conservateur: Sodium azide

Précaution d'utilisation: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Stock: 4 °C

Stockage commentaire: Store undiluted at 4°C.

Références pour anticorps anti-IL-8 (ABIN2689785)

Matsushima, Oppenheim: "Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF." dans: Cytokine, Vol. 1, Issue 1, pp. 2-13, (1991) (PubMed).

Hsu, Raine, Fanger: "Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures." dans: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 29, Issue 4, pp. 577-80, (1981) (PubMed).

Détails sur IL-8

Antigène: IL-8 (IL8) (Interleukin 8 (IL8))

Autre désignation: IL-8 (IL8 Produits)

Synonymes: anticorps CXCL8, anticorps GCP-1, anticorps GCP1, anticorps LECT, anticorps LUCT, anticorps LYNAP, anticorps MDNCF, anticorps MONAP, anticorps NAF, anticorps NAP-1, anticorps NAP1, anticorps IL-8, anticorps AMCF-I, anticorps il8, anticorps IL8, anticorps si:dkey-151b16.2, anticorps cxcli2, anticorps cxcl8, anticorps gcp-1, anticorps il-8, anticorps lynap, anticorps mdncf, anticorps monap, anticorps nap-1, anticorps C-X-C motif chemokine ligand 8, anticorps interleukin 8, anticorps chemokine (C-X-C motif) ligand 8a, anticorps chemokine (C-X-C motif) ligand 1-like, anticorps chemokine (C-X-C motif) ligand 8 L homeolog, anticorps CXCL8, anticorps il8, anticorps cxcl8a, anticorps IL8, anticorps Cxcl8, anticorps LOC422654, anticorps cxcl8.L

Pathways: Signalisation TLR, Cellular Response to Molecule of Bacterial Origin, Regulation of G-Protein Coupled Receptor Protein Signaling, ER-Nucleus Signaling, Hepatitis C, Autophagy